利用马铃薯X病毒表达载体在植物中表达非洲猪瘟病毒p30蛋白

In planta expression of African swine fever virus(ASFV)p30 protein using a potato virus X-based expression vector

由非洲猪瘟病毒(African swine fever virus,ASFV)引起的非洲猪瘟疫病(ASF)具有极高发病率和致死率,是我国一类动物疫病,对我国养猪产业造成重大威胁和巨大经济损失。p30蛋白是ASFV最主要结构蛋白之一,具有强烈免疫原性,是ASFV疫苗研发重要靶标。利用马铃薯X病毒(Potato virus X,PVX)侵染性克隆,构建表达p30蛋白重组PVX载体,通过农杆菌浸润法验证在模式植物本氏烟中表达p30蛋白可行性。结果表明,重组PVX载体接种本氏烟后5 d,植株系统叶片出现花叶症状;接种11 d后,植株系统叶片明显坏死。RT-PCR和Western blot检测表明p30蛋白在本氏烟中成功表达。研究结果为进一步规模化生产植物源p30蛋白奠定坚实基础。

African swine fever(ASF),an infectious disease caused by African swine fever virus(ASFV)with high incidence and mortality,is listed in the animal epidemics of class I in China,and has caused heavy economic losses to the pig breeding industry of China.The p30 protein,one of the major structural proteins of ASFV with strong antigenicity,is an important target for developing ASFV vaccine.In this study,a recombinant p30-expressing vector was constructed using the infectious clone of potato virus X(PVX)and was used to infect the model plant Nicotiana benthamiana by agroinfiltration for expressing p30 in plants.The systemic leaves of N.benthamiana inoculated by the recombinant vector showed mosaic symptoms at 5 days post-inoculation(dpi),and necrosis at 11 dpi.The results of RT-PCR and Western blot suggested that p30 protein had been successfully expressed in N.benthamiana.These results established a solid base for large-scale production of p30 protein in plants.